The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.

Respiratory syncytial virus (RSV) is an RNA virus in the Family Paramyxoviridae. Here, the activities performed by the RSV polymerase when it encounters the viral antigenomic promoter were examined. RSV RNA synthesis was reconstituted in vitro using recombinant, isolated polymerase and an RNA oligon...

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Main Authors: Sarah L Noton, Laure R Deflubé, Chadene Z Tremaglio, Rachel Fearns
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS Pathogens
Online Access:http://europepmc.org/articles/PMC3475672?pdf=render
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author Sarah L Noton
Laure R Deflubé
Chadene Z Tremaglio
Rachel Fearns
author_facet Sarah L Noton
Laure R Deflubé
Chadene Z Tremaglio
Rachel Fearns
author_sort Sarah L Noton
collection DOAJ
description Respiratory syncytial virus (RSV) is an RNA virus in the Family Paramyxoviridae. Here, the activities performed by the RSV polymerase when it encounters the viral antigenomic promoter were examined. RSV RNA synthesis was reconstituted in vitro using recombinant, isolated polymerase and an RNA oligonucleotide template representing nucleotides 1-25 of the trailer complement (TrC) promoter. The RSV polymerase was found to have two RNA synthesis activities, initiating RNA synthesis from the +3 site on the promoter, and adding a specific sequence of nucleotides to the 3' end of the TrC RNA using a back-priming mechanism. Examination of viral RNA isolated from RSV infected cells identified RNAs initiated at the +3 site on the TrC promoter, in addition to the expected +1 site, and showed that a significant proportion of antigenome RNAs contained specific nucleotide additions at the 3' end, demonstrating that the observations made in vitro reflected events that occur during RSV infection. Analysis of the impact of the 3' terminal extension on promoter activity indicated that it can inhibit RNA synthesis initiation. These findings indicate that RSV polymerase-promoter interactions are more complex than previously thought and suggest that there might be sophisticated mechanisms for regulating promoter activity during infection.
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spelling doaj.art-f19f1f903efd4d5a8c40fd183725bc5b2022-12-22T01:31:38ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742012-01-01810e100298010.1371/journal.ppat.1002980The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.Sarah L NotonLaure R DeflubéChadene Z TremaglioRachel FearnsRespiratory syncytial virus (RSV) is an RNA virus in the Family Paramyxoviridae. Here, the activities performed by the RSV polymerase when it encounters the viral antigenomic promoter were examined. RSV RNA synthesis was reconstituted in vitro using recombinant, isolated polymerase and an RNA oligonucleotide template representing nucleotides 1-25 of the trailer complement (TrC) promoter. The RSV polymerase was found to have two RNA synthesis activities, initiating RNA synthesis from the +3 site on the promoter, and adding a specific sequence of nucleotides to the 3' end of the TrC RNA using a back-priming mechanism. Examination of viral RNA isolated from RSV infected cells identified RNAs initiated at the +3 site on the TrC promoter, in addition to the expected +1 site, and showed that a significant proportion of antigenome RNAs contained specific nucleotide additions at the 3' end, demonstrating that the observations made in vitro reflected events that occur during RSV infection. Analysis of the impact of the 3' terminal extension on promoter activity indicated that it can inhibit RNA synthesis initiation. These findings indicate that RSV polymerase-promoter interactions are more complex than previously thought and suggest that there might be sophisticated mechanisms for regulating promoter activity during infection.http://europepmc.org/articles/PMC3475672?pdf=render
spellingShingle Sarah L Noton
Laure R Deflubé
Chadene Z Tremaglio
Rachel Fearns
The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
PLoS Pathogens
title The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
title_full The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
title_fullStr The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
title_full_unstemmed The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
title_short The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
title_sort respiratory syncytial virus polymerase has multiple rna synthesis activities at the promoter
url http://europepmc.org/articles/PMC3475672?pdf=render
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